LOCUS       sequence-263815-        6920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP-DAXX fusion protein with a mutation in its 5' SIM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6920)
  AUTHORS   Shastrula PK, Sierra I, Deng Z, Keeney F, Hayden JE, Lieberman PM, 
            Janicki SM
  TITLE     PML is recruited to heterochromatin during S phase and represses 
            DAXX-mediated histone H3.3 chromatin assembly.
  JOURNAL   J Cell Sci. 2019 Mar 26;132(6). pii: jcs.220970. doi: 
            10.1242/jcs.220970.
  PUBMED    30796101
REFERENCE   2  (bases 1 to 6920)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2019-03-26"; volume: "132(6). pii: jcs.220970. doi"; 
            pages: " 10.1242/jcs.220970"
COMMENT     Sequence Label: YFP-DAXX-mSIM1
FEATURES             Location/Qualifiers
     source          1..6920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3344..3465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3381..3400)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3435..3454
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3472..3927)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3609..3630)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3821..3840
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3954..4058
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4060..4417
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4060..4251
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4083..4103)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4268..4403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4452..5246
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4452..5246
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4506..4525)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5116..5135
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5434..5453)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5478..5525
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5854..6442
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6343..6362
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6617..6920
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ttcggctacg gcctgaagtg cttcgcccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcatg gacgagctgt
      961 acaagtactc agatctcgag atggccaccg ctaacagcat cggcgcgggg gatgatgatg
     1021 acgaagatga agcagctgct cagccagggc cctcccaccc actccccaat gcggcctcac
     1081 ctggggcaga agcccctagc tcctctgagc ctcatggggc cagaggaagc agtagttcgg
     1141 gcggcaagaa atgctacaag ctggagaatg agaagctgtt cgaagagttc cttgaacttt
     1201 gtaagatgca gacagcagac caccctgagg tggtcccatt cctctataac cggcagcaac
     1261 gtgcccactc tctgtttttg gcctcggcgg agttctgcaa catcctctct agggtcctgt
     1321 ctcgggcccg gagccggcca gccaagctct atgtctacat caatgagctc tgcactgttc
     1381 tcaaggccca ctcagccaaa aagaagctga acttggcccc tgccgccacc acctccaatg
     1441 agccctctgg gaataaccct cccacacacc tctccttgga ccccacaaat gctgaaaaca
     1501 ctgcctctca gtctccaagg acccgtggtt cccggcggca gatccagcgt ttggagcagc
     1561 tgctggcgct ctatgtggca gagatccggc ggctgcagga aaaggagtta gatctgagcg
     1621 aattggatga cccagactcc gcatacctgc aggaggcacg gttgaagcgt aagctgatcc
     1681 gcctctttgg gcgactatgt gagctgaaag actgctcttc actgaccggc cgtgtcatag
     1741 agcagcgcat cccctaccgt ggcacccgct acccagaggt taacaggcgc attgagcggc
     1801 tcatcaacaa gccagggcct gataccttcc ctgactatgg ggatgtgctt cgggctgtag
     1861 agaaggcagc tgcccgacac agccttggcc tcccccgaca gcagctccag ctcatggctc
     1921 aggatgcctt ccgagatgtg ggcatcaggt tacaggagcg acgtcacctc gatctcatct
     1981 acaactttgg ctgccacctc acagatgact ataggccagg cgttgaccct gcactatcag
     2041 atcctgtgtt ggcccggcgc cttcgggaaa accggagttt ggccatgagt cggctggatg
     2101 aggtcatctc caaatacgca atgttgcaag acaaaagtga ggagggcgag agaaaaaaga
     2161 gaagagctcg gctccaaggc acctcttccc actctgcaga cacccccgaa gcctccttgg
     2221 attctggtga gggccctagt ggaatggcat cccaggggtg cccttctgcc tccagagctg
     2281 agacagatga cgaagacgat gaggagagtg atgaggaaga ggaggaggag gaggaagaag
     2341 aagaggagga ggccacagat tctgaagagg aggaggatct ggaacagatg caggagggtc
     2401 aggaggatga tgaagaggag gacgaagagg aagaagcagc agcaggtaaa gatggagaca
     2461 agagccccat gtcctcacta cagatctcca atgaaaagaa cctggaacct ggcaaacaga
     2521 tcagcagatc ttcaggggag cagcaaaaca aaggacgcat agtgtcacca tcgttactgt
     2581 cagaagaacc cctggccccc tccagcatag atgctgaaag caatggagaa cagcctgagg
     2641 agctgaccct ggaggaagaa agccctgtgt ctcagctctt tgagctagag attgaagctt
     2701 tgcccctgga taccccttcc tctgtggaga cggacatttc ctcttccagg aagcaatcag
     2761 aggagccctt caccactgtc ttagagaatg gagcaggcat ggtctcttct acttccttca
     2821 atggaggcgt ctctcctcac aactggggag attctggtcc cccctgcaaa aaatctcgga
     2881 aggagaagaa gcaaacagga tcagggccat taggaaacag ctatgtggaa aggcaaaggt
     2941 cagtgcatga gaagaatggg aaaaagatat gtaccctgcc cagcccacct tcccccttgg
     3001 cttccttggc cccagttgct gattcctcca cgagggtgga ctctcccagc catggcctgg
     3061 tgaccagctc cctctgcatc ccttctccag cccggctgtc ccaaaccccc cattcacagc
     3121 ctcctcggcc tggtacttgc aagacaagtg tggccacaca atgcgatcca gaagagatca
     3181 tcgtgctctc agactctgat tagccgcggg cccgggatcc accggatcta gataactgat
     3241 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3301 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3361 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     3481 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3541 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3601 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3661 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3721 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3781 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3841 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3901 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3961 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4021 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4081 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4141 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4201 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4261 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4321 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4381 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4441 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4501 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4561 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4621 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4681 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4741 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4801 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4861 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4921 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4981 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5041 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5101 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5161 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5221 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5281 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5341 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5401 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     5461 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     5521 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     5581 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5641 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5701 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5761 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5821 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5881 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5941 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6001 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6061 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6121 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6181 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacaacg
     6241 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6301 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6361 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6421 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6481 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6541 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6601 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     6661 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     6721 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     6781 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     6841 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     6901 tagtcatcgc tattaccatg
//
